Access
To read this article in full you may need to log in, make a payment or gain access through a site license (see right).
Protocol
Nature Protocols 1, 1213–1222 (1 September 2006) | doi:10.1038/nprot.2006.127
Expanded-bed adsorption immobilized-metal affinity chromatography
&
Abstract
The protocol describes a method for capture of secreted hexahistidine-tagged proteins using expanded-bed adsorption immobilized-metal affinity chromatography. The starting material for the procedure is any crude feedstock that contains a histidine (His)-tagged target protein. The protocol is exemplified using unclarified broth from Pichia pastoris fermentation as feedstock. The protocol can be used for laboratory studies or as part of a process for production of recombinant biotherapeutics to standards of good manufacturing practice. It takes approximately 5 h to purify proteins from 10 liters of feedstock and a further 5–6 h to sterilize and regenerate the column.
To read this article in full you may need to log in, make a payment or gain access through a site license (see right).
